It is proposed to characterize active factors operating in recently developed methods for the in vitro culture and activation of pluripotential hemopoietic cells and to transfer DNA's for GM- CSF and MultiCSF into such cultured stem cells or their controlling stromal cells to determine the role of dysregulated CSF production in the development and progression of myeloid leukemia. The biochemical nature of the factors able to control the maintenance and expansion of pluripotential hemopoietic cell numbers will be determined using three types of starting materials (a) medium from the cloned B.Ad stromal cell line, (b) normal organ conditioned media (thymus, marrow shaft), and (c) U5637 conditioned medium. Biochemical fractionation will be monitored by three assay systems using post5FU bone marrow cells and will determine whether more than one such factor exists. Where appropriate, FACSpurified pluripotential cells will be used with purified recombinant CSF's to establish whether the stem cell-active factors have proliferative, activating or differentiation-inducing actions. DNA's for murine GM-CSF and Multi-CSF (IL-3) will be introduced (a) into hemopoietic stemm cells cultured with stimulation by the above factors, sing retroviral vectors and microinjection, then the cells used to repopulate irradiated recipients, and (b) into all tissues of the body by the generation of transgenic mice. The hemopoietic tissues from these contrasting mice will be analyzed to determine whether dysregulated CSF synthesis alone is sufficient for myeloid leukemia development or whether additional radiationinduced translations or the insertion of an additional oncgene are necessary.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA025972-08
Application #
3167123
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1983-06-01
Project End
1989-11-30
Budget Start
1987-12-01
Budget End
1988-11-30
Support Year
8
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Walter and Eliza Hall Institute Medical Research
Department
Type
DUNS #
City
Victoria
State
Country
Australia
Zip Code
3050
McArthur, G A; Metcalf, D; Rakar, S et al. (1995) Overexpression of C-FMS in the myeloid cell line FDC-P1 induces transformation that dissociates M-CSF-induced proliferation and differentiation. Leukemia 9:68-76
Li, C L; Johnson, G R (1995) Murine hematopoietic stem and progenitor cells: I. Enrichment and biologic characterization. Blood 85:1472-9
McArthur, G A; Longmore, G D; Klingler, K et al. (1995) Lineage-restricted recruitment of immature hematopoietic progenitor cells in response to Epo after normal hematopoietic cell transfection with EpoR. Exp Hematol 23:645-54
Li, C L; Wu, L; Antica, M et al. (1995) Purified murine long-term in vivo hematopoietic repopulating cells are not prothymocytes. Exp Hematol 23:21-5
Li, C L; Johnson, G R (1994) Stem cell factor enhances the survival but not the self-renewal of murine hematopoietic long-term repopulating cells. Blood 84:408-14
McArthur, G A; Rohrschneider, L R; Johnson, G R (1994) Induced expression of c-fms in normal hematopoietic cells shows evidence for both conservation and lineage restriction of signal transduction in response to macrophage colony-stimulating factor. Blood 83:972-81
Duhrsen, U; Novotny, J; Boyd, A W (1994) Self-renewal of a transplantable murine leukemia induced by co-culture with human stromal cell lines. Leukemia 8:490-7
Li, C L; Johnson, G R (1992) Long-term hemopoietic repopulation by Thy-1lo, Lin-, Ly6A/E+ cells. Exp Hematol 20:1309-15
Metcalf, D; Nicola, N A; Gough, N M et al. (1992) Synergistic suppression: anomalous inhibition of the proliferation of factor-dependent hemopoietic cells by combination of two colony-stimulating factors. Proc Natl Acad Sci U S A 89:2819-23
Metcalf, D (1992) Mechanisms responsible for size differences between hemopoietic colonies: an analysis using a CSF-dependent hemopoietic cell line. Int J Cell Cloning 10:116-25

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